Information on EC 3.2.1.75 - glucan endo-1,6-beta-glucosidase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
3.2.1.75
-
RECOMMENDED NAME
GeneOntology No.
glucan endo-1,6-beta-glucosidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Random hydrolysis of (1->6)-linkages in (1->6)-beta-D-glucans
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of O-glycosyl bond
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
6-beta-D-glucan glucanohydrolase
Acts on lutean, pustulan and 1,6-oligo-beta-D-glucosides.
CAS REGISTRY NUMBER
COMMENTARY hide
37278-39-0
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
4 different species
-
-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
WL-12
-
-
Manually annotated by BRENDA team
WL-12
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Flavobacterium M64
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Fusarium roseum
-
-
-
Manually annotated by BRENDA team
Helminthosporium sesanum
-
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Neotyphodium sp.
-
SwissProt
Manually annotated by BRENDA team
strain IFO-6068
-
-
Manually annotated by BRENDA team
QM 1873
-
-
Manually annotated by BRENDA team
QM 1873
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
10 different species
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
NCYC 451
-
-
Manually annotated by BRENDA team
NCYC 451
-
-
Manually annotated by BRENDA team
growth medium with lyophilized cells of Candida utilis as mian nutrient source
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-
Manually annotated by BRENDA team
CECT 2313
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-
Manually annotated by BRENDA team
strain CECT 2413, 3 isozymes BGN16.1, BGN16.2, and BGN16.3
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-
Manually annotated by BRENDA team
several strains: A.11.178, A.9.3., B.1.2, T.16.C, T.2.C
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-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
4-nitrophenyl beta-gentiohexaoside + H2O
?
show the reaction diagram
4-nitrophenyl beta-gentiopentaoside + H2O
?
show the reaction diagram
-
-
-
-
?
4-nitrophenyl beta-gentiotetraoside + H2O
?
show the reaction diagram
-
-
-
-
?
amylopectin + H2O
?
show the reaction diagram
-
-
-
-
?
amylose + H2O
?
show the reaction diagram
beta-(1,6)-glucan + H2O
?
show the reaction diagram
-
highly branched beta-(1,6)-glucan, that contains about 75-80% of linear beta-(1,6)-glucose linked units, preared from yeast cell walls
-
-
?
beta-glucan + H2O
glucose + laminari-oligosaccharides + gentio-oligosaccharides
show the reaction diagram
carboxymethyl cellulose + H2O
?
show the reaction diagram
carboxymethyl-pachyman + H2O
?
show the reaction diagram
carboxymethylcellulose + H2O
?
show the reaction diagram
-
-
-
-
?
cellulose + H2O
?
show the reaction diagram
chitosan + H2O
?
show the reaction diagram
-
3% of the activity with pustulan
-
-
?
curdlan + H2O
?
show the reaction diagram
-
0.3% of the activity against pustulan
-
-
?
dextran + H2O
?
show the reaction diagram
Eisenia bicyclis laminarin + H2O
?
show the reaction diagram
27% of the activity with pustulan
-
-
?
gentiobiose + H2O
D-glucose
show the reaction diagram
gentiohexaose + H2O
gentiobiose + gentiotriose + gentiotetraose
show the reaction diagram
gentiooctaose + H2O
?
show the reaction diagram
gentiopentaose + H2O
D-glucose + gentiotetraose + gentiotriose + gentiobiose
show the reaction diagram
gentiopentaosylsorbitol + H2O
gentiotetraose + gentiobiose + D-glucose
show the reaction diagram
-
-
-
?
gentiotetraose + H2O
D-glucose + gentiotriose + gentiobiose
show the reaction diagram
gentiotriose + H2O
D-glucose + gentiobiose
show the reaction diagram
-
-
-
?
gentiotriosylsorbitol + H2O
?
show the reaction diagram
-
-
-
-
?
glucan + H2O
?
show the reaction diagram
glycogen + H2O
?
show the reaction diagram
gyrophoran + H2O
?
show the reaction diagram
-
-
-
-
?
islandican + H2O
?
show the reaction diagram
-
-
-
-
?
lactose + H2O
D-glucose
show the reaction diagram
-
-
-
?
laminaran + H2O
?
show the reaction diagram
laminarin + H2O
?
show the reaction diagram
laminarin + H2O
D-glucose + ?
show the reaction diagram
laminarin + H2O
gentiobiose + D-glucose
show the reaction diagram
lichenan + H2O
?
show the reaction diagram
lutean + H2O
?
show the reaction diagram
73% of the activity with pustulan
-
-
?
lutean + H2O
D-glucose + gentiobiose + gentiotriose
show the reaction diagram
luteose + H2O
gentiobiose + gentiotriose + gentiotetraose
show the reaction diagram
methyl beta-gentiobioside + H2O
methyl beta-D-glucoside + beta-D-glucose
show the reaction diagram
-
low activity
-
-
?
methyl beta-gentiohexaoside + H2O
methyl beta-D-glucoside + 2 beta-D-glucose + gentiobioside
show the reaction diagram
-
the major cleavages occur at bonds 2, 3, and 4 in approximately equal proportion, the nonreducing end glucosyl residues of Gen4-6 beta-OMe are either not or scarcely cleaved by the enzyme
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-
?
methyl beta-gentiopentaoside + H2O
methyl beta-D-glucoside + 3 beta-D-glucose
show the reaction diagram
-
the major cleavages occur at bonds 2, 3, and 4 in approximately equal proportion
-
-
?
methyl beta-gentiotetraoside + H2O
methyl beta-D-glucoside + 2 beta-D-glucose
show the reaction diagram
-
cleavage at the second (bond 2) and third (bond 3) glucosidic bonds from the methyl group
-
-
?
methyl beta-gentiotrioside + H2O
methyl beta-D-glucoside + gentiobioside
show the reaction diagram
-
the second glucosidic bond (bond 2) from the methyl group is cut preferentially
Glc beta-OMe and Gen2 beta-OMe in the ratio 91:9
-
?
p-nitrophenyl-beta-D-glucoside + H2O
4-nitrophenol + beta-D-glucose
show the reaction diagram
pachyman + H2O
gentiobiose
show the reaction diagram
pustulan + H2O
?
show the reaction diagram
pustulan + H2O
D-glucose + gentobiose + ?
show the reaction diagram
-
-
main products
-
?
pustulan + H2O
gentobiose + D-glucose + ?
show the reaction diagram
-
-
-
-
?
pustulan + H2O
glucose + gentiobiose
show the reaction diagram
pustulan + H2O
oligomers of pustulan
show the reaction diagram
starch + H2O
?
show the reaction diagram
succinoglycan + H2O
?
show the reaction diagram
Flavobacterium M64
-
-
-
-
?
xylan + H2O
?
show the reaction diagram
-
12% of the activity with pustulan
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
gentiooctaose + H2O
?
show the reaction diagram
pustulan + H2O
?
show the reaction diagram
additional information
?
-
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-Hydroxy-5-nitrobenzyl bromide
-
-
Dicyclohexylcarbodiimide
-
-
glucono-delta-lactone
-
-
N-Acetylimidazole
-
-
N-bromosuccinimide
-
-
N-ethyl-5-phenylisoxazolium 3'-sulfonate
-
-
p-hydroxymercuribenzoate
-
-
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
-
BGN16.3 is induced rapidly and strongly by fungal cell walls
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.014
Gentiotetraose
-
-
0.00012
lutean
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assumed molecular weight for lutean 650000 Da
-
0.0753
pustulan
-
assumed molecular weight for pustulan 17000 Da, or KM of 1.28 mg/ml
-
additional information
additional information
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.22
Flavobacterium M64
-
-
0.48
Neotyphodium sp.
-
5
-
intracellular enzyme
75.4
-
extracellular enzyme
83.3
-
pH 5.0, 30C
164.1
-
purified native enzyme, pH 4.0, 50C
188
-
purified enzyme
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.2 - 4.6
-
-
4.8
-
assay at
5 - 5.5
5 - 5.5
6
-
in Tris/succinate buffer
additional information
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 7
-
20% of maximal activity at pH 4.0 and pH 7.0
4.5 - 6.5
-
at least 80% of maximum activity in this range
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.1
-
acidic chromatofocusing
5.5
-
isoelectric focusing
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20000 - 25000
-
gel filtration
20000
-
gel filtration
25000 - 30000
-
gel filtration
30500
-
sedimentation equilibrium
37000
-
gel filtration
43000
-
extracellular enzyme, gel filtration
44300
-
gel filtration
51000
-
intracellular enzyme, gel filtration
54000
-
SDS-PAGE, gel filtration
55000
-
PAGE
65000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
-
1 * 65000, SDS-PAGE
monomer or dimer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
no glycoprotein
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2 - 9
-
purified native enzyme, 4C, 24 h, over 90% activity remaining
731583
3 - 7
-
-
136854
4 - 8
the enzyme is stable over a broad pH range from pH 4.0 to 8.0 when treated at 4C for 20 h
713912
4.5 - 9.5
-
-
136862
4.9 - 5.5
-
-
136849
5 - 8
-
-
136847
5 - 6
-
at -20C
136850
5 - 8
-
-
136850
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
-
stable for a long time at 30C, activity decrease at 40C
30 - 60
-
-
45
-
unstable above, presence of pustulan stabilizes
75
30 min, about 20% residual activity
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, pH6, 2 months, no loss of activity
-
-20C, repeated freezing and thwaing severely damages enzyme activity
-
-25C, 1 month, less than 10% loss of activity
-
2C, 1 week, 20% loss of activity
-
4C, 4 months, no loss of activity, -70C, 12 months, no loss of activity
-
4C, 6 months, no loss of activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
140fold
Flavobacterium M64
-
154fold, 68fold expressed in Pichia pastoris
-
486.5fold
-
ammonium sulfate precipitation, Sepharose Q column chromatography, Superdex 75 gel filtration, and DEAE-Toyopearly column chromatography
FLAG-tagged Neg1 protein purified by batch adsorption
-
intracellular enzyme 100fold, extracellular enzyme 75fold
-
native enzyme 244.3fold from culture supernatant by ammonium sulfate fractionation, cation exchange and affinity chromatography using gentiotetraose as ligand
-
native extracellular BGN16.3 26fold to homogeneity by beta-1,6-glycan affinity chromatography, chromatofocusing, and gel filtration
-
partial
Neotyphodium sp.
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
-
Neotyphodium sp.
expression in Pichia pastoris
-
expression in Saccharomyces cerevisiae
-
expression profile of BGN16.3
-
functional expression from yeast expression vector pYESH2 in Saccharomyces cerevisiae strain INVSc1. The recombinant enzyme does not show antifungal activity in yeast
-
into expression/secretion vector pINA1296+FLAG, expressed in Yarrowia lipolytica strain Po1g: MatA, leu2-270, ura3-302::pBR322-URA3, xpr2-322, axp-2, which contains an integrated pBR322 plasmid as a docking platform
-
recombinant expression in Pichia pastoris
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
the transcript level of endo-beta-1,6-glucanase Pus30A gene (lepus30a) is significantly increased in fruiting bodies 4 days after harvesting (approximately 18times in gills, 27times in stipes, and 32times in pilei) compared to fresh fruiting bodies
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
-
the enzyme is used for analysis of yeast polysaccharides cell wall composition in a method combines acid and enzymatic hydrolysis, and involves a chitinase from Streptomyces griseus, an endo/exo-beta-(1,3)-glucanase from Trichoderma species, recombinant endo-beta-(1,6)-glucanase from Trichoderma harzianum, and a beta-glucosidase from Aspergillus niger, method development and evaluation, overview
biotechnology